Design and Testing of a Pulsatile Conditioning System for Dynamic Endothelialization of Polyphenol-Stabilized Tissue Engineered Heart Valves.
Design and Testing of a Pulsatile Conditioning System for Dynamic Endothelialization of Polyphenol-Stabilized Tissue Engineered Heart Valves.
复制标题
DOI：
10.1007/s13239-010-0014-6
复制
发表时间：
2010-06
影响因子：
1.8
通讯作者：
中科院分区：
文献类型：
--
作者：
心脏瓣膜组织工程需要生物相容性和血液相容性支架，其经历重塑和再增殖，但也能在植入后立即承受苛刻的机械力。我们假设，可逆稳定的脱细胞猪瓣膜，接种内皮细胞和条件脉动生物反应器将铺平道路，为下一代组织工程心脏瓣膜（TEHV）。首先设计、制造和测试了一种新型阀门调节系统，以充分评估TEHV。生物反应器可正确关闭和打开阀门，并允许在无菌条件下采用多种安装方法。采用化学法对猪主动脉瓣根部进行脱细胞处理，并用五没食子酰葡萄糖（PGG）进行稳定化处理。通过测试对胶原酶和弹性蛋白酶的抗性、双轴力学分析和热变性概况来评估新型支架的性质。将猪主动脉内皮细胞接种到瓣叶上，并在生理肺动脉瓣压力下将整个主动脉根部固定在动态脉动心脏瓣膜生物反应器系统内，并在17天后分析细胞活力、形态和代谢活性。我们的组织制备方法有效地去除了细胞，包括有效的α-Gal抗原，同时保留了具有足够机械性能的保存良好的细胞外基质支架。PGG增强了细胞外基质成分的稳定性，但也显示出可逆的能力。工程瓣膜支架促进内皮细胞的附着和存活延长时间，并显示出调节后广泛细胞覆盖的迹象。我们的新方法显示出开发能够重塑和细胞再生的坚固耐用的TEHV的希望。
Heart valve tissue engineering requires biocompatible and hemocompatible scaffolds that undergo remodeling and repopulation, but that also withstand harsh mechanical forces immediately following implantation. We hypothesized that reversibly stabilized acellular porcine valves, seeded with endothelial cells and conditioned in pulsatile bioreactors would pave the way for next generations of tissue engineered heart valves (TEHVs). A novel valve conditioning system was first designed, manufactured and tested to adequately assess TEHVs. The bioreactor created proper closing and opening of valves and allowed for multiple mounting methods in sterile conditions. Porcine aortic heart valve roots were decellularized by chemical extractions and treated with penta-galloyl glucose (PGG) for stabilization. Properties of the novel scaffolds were evaluated by testing resistance to collagenase and elastase, biaxial mechanical analysis, and thermal denaturation profiles. Porcine aortic endothelial cells were seeded onto the leaflets and whole aortic roots were mounted within the dynamic pulsatile heart valve bioreactor system under physiologic pulmonary valve pressures and analyzed after 17 days for cell viability, morphology, and metabolic activity. Our tissue preparation methods effectively removed cells, including the potent α-Gal antigen, while leaving a well preserved extra-cellular matrix scaffold with adequate mechanical properties. PGG enhanced stabilization of extracellular matrix components but also showed the ability to be reversible. Engineered valve scaffolds encouraged attachment and survival of endothelial cells for extended periods and showed signs of widespread cell coverage after conditioning. Our novel approach shows promise toward development of sturdy and durable TEHVs capable of remodeling and cellular repopulation.